Screening differentially expressed genes in an amphipod (Hyalella azteca) exposed to fungicide vinclozolin by suppression subtractive hybridization

Vinclozolin, a dicarboximide fungicide, is an endocrine disrupting chemical that competes with an androgenic endocrine disruptor compound. Most research has focused on the epigenetic effect of vinclozolin in humans. In terms of ecotoxicology, understanding the effect of vinclozolin on non-target organisms is important. The expression profile of a comprehensive set of genes in the amphipod Hyalella azteca exposed to vinclozolin was examined. The expressed sequence tags in low-dose vinclozolin-treated and -untreated amphipods were isolated and identified by suppression subtractive hybridization. DNA dot blotting was used to confirm the results and establish a subtracted cDNA library for comparing all differentially expressed sequences with and without vinclozolin treatment. In total, 494 differentially expressed genes, including hemocyanin, heatshock protein, cytochrome, cytochrome oxidase and NADH dehydrogenase were detected. Hemocyanin was the most abundant gene. DNA dot blotting revealed 55 genes with significant differential expression. These genes included larval serum protein 1 alpha, E3 ubiquitin-protein ligase, mitochondrial cytochrome c oxidase, mitochondrial protein, proteasome inhibitor, hemocyanin, zinc-finger–containing protein, mitochondrial NADH-ubiquinone oxidoreductase and epididymal sperm-binding protein. Vinclozolin appears to upregulate stress-related genes and hemocyanin, related to immunity. Moreover, vinclozolin downregulated NADH dehydrogenase, related to respiration. Thus, even a non-lethal concentration of vinclozolin still has an effect at the genetic level in H. azteca and presents a potential risk, especially as it would affect non-target organism hormone metabolism.     

Temperature dependence of the absolute rate constant for the reaction of ozone with dimethyl sulfide

Absolute rate constants for the reaction of ozone with dimethyl sulfide (DMS) were measured in a 200-L Teflon chamber over the temperature range of 283-353 K. Measurements were carried out using DMS in large excess over ozone of 10 to 1 or greater. Over the indicated temperature range,the data could be fit to the simple Arrhenius expression as KDMS = (9.96±3.61)×10-11exp(-(7309.7±1098.2)/T)cm3/(molecule·s). A compared investigation of the reaction between ozone and ethene had a kc2H4 value of(1.35±0.11)×10-18 cm3/(molecule·s) at room temperature.     

2种烷基多环芳烃对仿刺参CYP450和p53基因表达的影响研究

海洋环境中多环芳烃类(PAHs)主要来源于海洋溢油事故以及沿海石油化工企业的废水排放,国内外大量研究发现海洋中的多环芳烃对海洋生物造成了潜在的生态风险。为了揭示不同浓度多环芳烃类污染物对海参的生态毒理效应,将仿刺参(Apostichopus japonicus)分别暴露于不同浓度的2种烷基多环芳烃3-甲基菲(5、10、100μg·L~(-1))和2-甲基蒽(5、10、50μg·L~(-1))中,检测暴露3 d、7 d和14 d后,3-甲基菲和2-甲基蒽胁迫下仿刺参CYP450和p53基因的相对表达量。结果表明,3-甲基菲和2-甲基蒽胁迫下,仿刺参CYP450和p53基因的表达均对毒物产生了不同程度的响应。与对照组相比,3-甲基菲各处理组对仿刺参CYP450和p53基因的表达均产生显著的抑制作用(P0.05); 2-甲基蒽各处理组对仿刺参CYP450和p53基因的表达影响作用不同,暴露7 d后,2-甲基蒽各处理组对仿刺参CYP450基因的表达表现出抑制作用,对p53基因的表达表现出诱导作用。相同浓度与时间胁迫下,2-甲基蒽对仿刺参CYP450和p53基因表达的影响比3-甲基菲的影响大。上述研究结果表明,3-甲基菲和2-甲基蒽均可不同程度影响仿刺参CYP450和p53基因的表达,且与3-甲基菲相比,2-甲基蒽对仿刺参CYP450和p53基因表达的影响较明显。上述结果为多环芳烃类污染物对仿刺参的生物毒性评价提供了基础数据。     

Biological treatment of chicken feather waste for improved biogas production

A two-stage system was developed which combines the biological degradation of keratin-rich waste with the production of biogas. Chicken feather waste was treated biologically with a recombinant Bacillus megaterium strain showing keratinase activity prior to biogas production. Chopped, autoclaved chicken feathers (4%, W/V) were completely degraded, resulting in a yellowish fermentation broth with a level of 0.51 mg/mL soluble proteins after 8 days of cultivation of the recombinant strain. During the subsequent anaerobic batch digestion experiments, methane production of 0.35 Nm³/kg dry feathers (i.e., 0.4 Nm³/kg volatile solids of feathers), corresponding to 80% of the theoretical value on proteins, was achieved from the feather hydrolyzates, independently of the pre-hydrolysis time period of 1, 2 or 8 days. Cultivation with a native keratinase producing strain, Bacillus licheniformis resulted in only 0.25 mg/mL soluble proteins in the feather hydrolyzate, which then was digested achieving a maximum accumulated methane production of 0.31 Nm³/kg dry feathers. Feather hydrolyzates treated with the wild type B. megaterium produced 0.21 Nm³ CH₄/kg dry feathers as maximum yield.     

氯化甲基汞对大鼠脑c-fos和c-jun基因表达的影响

为了研究即刻早期基因c-fos、c-jun在氯化甲基汞(MMC)神经毒性机制中的作用,本研究应用免疫组织化学方法对皮下注射甲基汞5.0mg/kg及0.1mg/kg的大鼠脑区FOS、JUN表达水平进行观察,对照组注射生理盐水.结果表明,急性暴露3h后,暴露组大鼠脑FOS、JUN蛋白表达均高于对照组,从而表明即刻早期基因(IEG)参与了氯化甲基汞对中枢神经系统损害的毒性过程.     

Estrogenic activities of two synthetic pyrethroids and their metabolites

Synthetic pyrethroids(SPs) are among the most common pesticides in current use,and so far,several SPs have been assessed for their potential estrogenicities by various methods.Previous studies have shown that the estrogenicities partly come from their metabolites.Although considerable information is available with respect to the metabolism and environmental degradation of SPs,little is known about the estrogenicities of the metabolites.In this study,permethrin(PM) and β-cypermethrin(CP),as well as their metabolites(3-phenoxybenzoic alcohol(PBCOH),3-phenoxybenzaldehyde(PBCHO) and 3-phenoxybenzoic acid(PBCOOH) were evaluated for their estrogenic activities in the MCF-7 human breast carcinoma cell line.In the MCF-7 cell proliferation assay,PM and CP exhibited significant estrogenic activities at 10-7 mol/L,comparable to 17β-estradiol(E2 ) of 10-9 mol/L,with the relative proliferative effect ratios of 55.4% and 56.3%,respectively.The real-time quantitative polymerase chain reaction(qRT-PCR) results confirmed the estrogenicities of PM and CP with significant alteration of pS2 and ERα mRNA levels observed at 10-6 mol/L.For the three major metabolites,PBCOH and PBCOOH exhibited estrogenic activities in all assays,while no significant estrogenic responses was observed for PBCHO compared to the vehicle control.In particular,PBCOH had even slightly stronger estrogenic activity than its parent compounds,indicating that metabolism may be one of the reasons for the estrogenicities of the SPs.Given the widespread use of SPs,the toxicological effects of parent compounds and their metabolites should be taken into consideration in the risk assessment of SPs.     

Prenatal diagnosis of severe combined immunodeficiency with defective synthesis of HLA molecules

The immunodeficiency associated with a defective expression of HLA molecules is an autosomal recessive disorder leading to death during childhood. We have performed prenatal diagnosis for six fetuses at risk for this disease by membrane immunofluorescence on blood lymphocytes and monocytes, using specific monoclonal antibodies for HLA class I and II molecules. Two pregnancies have been found to be affected. The diagnosis has been confirmed on each abortus by the study of the membrane expression of HLA class I and II molecules on blood lymphocytes and monocytes, and on thymic and splenic cells. The four other cases were found to be normal both during pregnancy and after birth. The detection of the defect as early as the 20th week of gestation allows selective termination.     

Prenatal testing for inherited immune deficiencies by fetal blood sampling

We attempted to develop a prenatal diagnosis in fetuses at risk for immunodeficiency by fetal blood sampling performed under fetoscopy at 18–22 weeks of gestation. In order to obtain normal values, we first investigated thirty-five control fetuses whose blood punctures were undertaken for the diagnosis of haemoglobinopathies. Surface markers and in vitro mitogen-induced proliferation of the fetal lymphocytes were studied using micromethods. We then examined two fetuses at risk for two different types of severe combined immunodeficiency and established their immunological integrity, hence avoiding an unjustified termination of pregnancy. This immunological integrity was confirmed after birth.     

围生期双酚A暴露对子代大鼠脾脏免疫功能的影响

为了研究围生期双酚A(BPA)暴露对子代脾脏免疫功能的影响,选择F344大鼠,按体重随机分为对照、低、中、高4个剂量组(BPA暴露剂量分别为0、4、40、400mg·kg-1·d-1),从雌鼠妊娠第0天开始至子鼠出生后30天,每天灌胃染毒1次,观察子鼠脾脏组织形态学的变化,并采用实时定量PCR检测IL-2、IL-12、IFN-γ、TNF-α4种细胞因子mRNA表达的改变.结果表明,1)与对照组相比,低剂量组子鼠脾重、脾脏指数均显著增加(p<0.05),体重无显著变化(p>0.05);中剂量组子鼠脾脏指数显著增加(p<0.05),体重和脾重无显著变化(p>0.05);高剂量组子鼠体重、脾重均显著降低(p<0.05),脾脏指数无显著变化(p>0.05).2)与对照组相比,各暴露组子鼠脾脏IL-2、IL-12、IFN-γ、TNF-α4种细胞因子mRNA表达均显著降低(p<0.01).以上结果提示,BPA可能能够透过胎盘屏障并通过乳汁传递给子代,降低子代的机体免疫功能.     

一种新去整合素Adinbitor的表达、序列分析及其对C6神经胶质瘤细胞凋亡的诱导

Adinbitor为一通过基因工程方法获得的来自于中国白眉蝮蛇毒腺的去整合素家族的新成员 .从蝮蛇毒腺中提取总RNA进行RT -PCR扩增 ,获得了 2 19bp的白眉蝮蛇去整合素基因 .cDNA测序及由此推导的氨基酸序列结果显示 ,Adinbitor为含有 73个氨基酸的多肽 ,其中包括 12个半胱氨酸 ,并具有去整合素的特征模序 -RGD .对此去整合素基因进行克隆、转化与诱导后 ,得到了该蛋白的可溶性高效表达 .经组氨酸亲和层析纯化 ,获得了分子量Mr为 9×10 3 的均质融合蛋白 ,并将其命名为adinbitor.序列分析表明 ,adinbitor与已发表于GENBANK上的整合素序列有很高的同源性 ,其中同源性最高的是南韩的saxatilin ,二者的蛋白质同源性高达 91.78% .活性测定结果表明 ,Adinbitor能有效地诱导神经胶质瘤细胞C6的细胞凋亡 ,有希望成为一种抗癌新药 .图 5参 15